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Advances in diagnostic tests for bacterial STDs.

Because of their asymptomatic nature and nonspecific symptoms, laboratory tests are often required to diagnose a sexually transmitted infection. Over the past few years, there have been advances in technology, such as the development of nucleic acid amplification assays, which have improved our ability to diagnose infections caused by Chlamydia trachomatis. The finding that nucleic acid amplification tests can detect more infected individuals and are useful in screening low prevalence populations, has led to the development of strategies designed to reduce the cost of these assays without significantly impacting their sensitivity. The development of new tests for the diagnosis of syphilis has gained momentum from the report of a synthetic VDRL antigen, which will result in better nontreponemal antibody tests for syphilis. In spite of the completion of the genome sequence of Treponema pallidum and its annotation, we are still unable to cultivate this microorganism in vitro. However, the molecular revolution has resulted in the development of PCR assays for detecting Treponema pallidum in various types of clinical specimens, and to the production of recombinant antigens for use in tests that detect treponemal-specific antibodies. Further research will improve the availability of low cost, sensitive tests for the diagnosis of sexually transmitted infections. The English version of this paper is available too at:http://www.insp.mx/salud/index.html.

Bacterial Infections↗

Role of fibronectin in the pathogenesis of syphilis.

In recent years studies concerned with the attachment of Treponema pallidum to host cells have focused on the possible, role of fibronectin in mediation of this event. Although our understanding of in vitro fibronectin-T. pallidum interactions has increased markedly, considerable controversy exists concerning not only the kinetics of binding but whether fibronectin is the only matrix protein involved in attachment. Moreover, a unifying hypothesis concerned with the role of fibronectin in the pathogenesis of syphilis cannot be limited solely to early events. Further characterization of fibronectin-T. pallidum interactions must also take into consideration that the natural infection, characterized by distinct stages, is accompanied by immunologic responses that undoubtedly impact on those cellular interactions.

Bacterial Adhesion↗

Administration of antithymocyte serum to syphilitic rabbits inhibits development of resistance to reinfection with Treponema pallidum.

Three groups of female rabbits were inoculated i.v. with T. pallidum. One group of the animals was treated with horse anti-rabbit thymocyte serum (ATS) one day before and during the next two weeks after infection. Two other groups, one treated with normal horse serum (NHS) in the same way as the previous one and the other untreated, served as controls. After various time intervals, all the animals were tested for the content of T and B lymphocytes in the peripheral blood, humoral (VDRL and TPI tests), and cell-mediated (SDH test) responses. Two days after testing, animals of each group were challenged i.d. with T. pallidum, and observed for appearance of syphilitic lesions. The treatment of syphilitic rabbits with ATS caused a significant reduction in the number of T lymphocytes and a distinct inhibition of development of cell-mediated immunity associated with the lack of resistance to reinfection with T. pallidum till the 16th week of the infection; the level of B lymphocytes and the development of serological responsiveness was not affected by ATS treatment. The results are interpreted as indicating the protective role of cell-mediated immunity in syphilis.

Animals↗

The time-dependent clearance of virulent Treponema pallidum in susceptible and resistant strains of guinea pigs is significantly different.

The kinetics of clearance of Treponema pallidum spp. pallidum Nichols from skin and testes of susceptible C4-deficient (C4D) and -resistant Albany (Alb) strains of guinea pigs (gps) was evaluated using the polymerase chain reaction (PCR) and the rabbit infectivity test (RIT). For each strain there were two groups of animals, one infected with virulent T. pallidum (TP) and one control injected with heat-killed treponemes (HKTP). The kinetic studies and their statistical analysis showed that in the C4D strain the microbial clearance in both tissues was significantly slower (p < 0.005) and still incomplete at 3 months after infection. In the Alb strain the clearance was faster and apparently completed within a month. A greater permissiveness in bacterial growth in C4D compared to Alb appears to be one critical factor determining the different rate of local elimination after primary infection. In both strains there was some correlation between the severity and duration of cutaneous lesions and the local persistence of viable organisms. This correlation was not observed in testes. These studies suggest a genetic basis for the strain-specific susceptibility and resistance phenotypes in the pathogenesis of syphilis.

Animals↗

Syphilitic uveitis as the initial manifestation of HIV infection.

Syphilis is an uncommon cause of uveitis in HIV-infected patients. We report a case of bilateral panuveitis and describe its characteristics as the initial manifestation of HIV infection. A 74-year-old heterosexual male complained of blurred vision and floaters in both eyes for 40 days. Slit lamp examination showed diffuse keratic precipitates and cells in the anterior chamber of both eyes. Fundus examination revealed multiple small white dots and scattered retinal hemorrhage over the mid-equatorial retina with marked vitritis. Physical examination disclosed multiple erythematous papules over bilateral palms compatible with secondary syphilis. Serologic tests--the venereal disease research laboratory (VDRL) test, fluorescent treponemal antibody absorption (FTA-ABS) test, and Treponema pallidum hemagglutination (TPHA) test--were all positive. Aqueous fluid also showed positive FTA-ABS reaction. Under the impression of acquired secondary syphilis, enzyme-linked immunosorbent assay and Western blot test were performed and revealed concurrent HIV infection. After intravenous administration of penicillin-G, 18 million units daily for 2 weeks, the vitritis and retinochoroiditis improved. All patients with panuveitis of unknown cause should undergo VDRL and FTA-ABS screening. Subsequent testing for HIV antibody in leutic uveitis is also mandatory.

Aged↗

Serological findings in leprosy. An investigation into the specificity of various serological tests for syphilis.

In serological tests for syphilis, leprosy sera often give biologically false positive reactions. These may be due to the presence of non-specific elements-for example, the ubiquitous lipid antibodies-in the leprosy sera; or they may be the result of errors in technique or unfavourable working conditions in the laboratory. This paper presents the results of an investigation in which several hundred sera from lepers were submitted to four of the so-called "standard" serological tests for syphilis (STS), using either cardiolipin or crude lipid antigens; to a complement-fixation test using as antigen a suspension of Reiter treponemes (PR test); and to the Treponema pallidum immobilization (TPI) test. The investigation was carried out in a moderate climate and in technically well-equipped laboratories.It was found that the number of biologically false positive reactions was not as high as had been expected in the light of previous investigations. It was discovered, moreover, that it was the lipid antigens that were mainly responsible for the non-specific reactions, since both the PR and the TPI test showed a far greater specificity than any of the STS. But the TPI test, though highly specific, is also technically very complicated and therefore not suitable for use in regions where technical facilities are lacking. The authors consider that, in such regions, the simpler PR test will give sufficiently accurate results in the serodiagnosis of treponematoses. It must, however, be recognized that even the treponemal tests are not capable of differentiating between syphilis and yaws infections.

Antibodies↗

Infectious causes of hydrops fetalis.

A variety of infectious agents have been associated with nonimmune hydrops fetalis, most notably parvovirus B19, cytomegalovirus, herpes simplex virus, Toxoplasma gondii, and Treponema pallidum. These agents produce hydrops through effects on fetal bone marrow, myocardium, or vascular endothelium. Knowledge of the epidemiology and clinical characteristics of maternal and fetal infection can be used to select a diagnostic approach. Etiologic diagnosis will guide prognosis and the selection of specific chemotherapy.

Female↗

Severe periodontitis in a 5-year-old girl with hyperimmunoglobulin E syndrome.

The hyperimmunoglobulin E syndrome (HIES) is a multisystem disorder that affects the: (1) dentition; (2) skeleton; (3) connective tissues; and (4) immune system. Little is known about periodontal manifestations of the syndrome. The purpose of this report was to describe a 5-year-old girl with suspected autosomal-recessive HIES syndrome who revealed profusely bleeding and painful gingiva and generalized aggressive periodontitis. A polymerase chain reaction (PCR)-based microbiological examination detected Porphyromonas gingivalis, Tannerella forsythia, Prevotella nigrescens, Treponema denticola, Eikenella corrodens, and Campylobacter rectus in the deep periodontitis lesions. The extraction of all deciduous teeth due to a poor prognosis and risk of systemic infection led to resolution of the oral inflammation. Long-term follow-up is required to determine the periodontal prognosis of the erupting permanent teeth.

Bacteria, Anaerobic↗

Infections among pygmies in the Eastern Province of Cameroon.

The health requirements of pygmies is poorly understood because of their continued isolation from the other tribes in Central Africa. This study was undertaken among the Baka pygmies of the Eastern Province of Cameroon to generate basic health data among them. A total of 141 adults (18-45 years) were tested for the hepatitis B surface antigen (HBsAg), antibody to the surface antigen (anti-HBs), antibody to the core antigen (anti-HBc) and antibody to the hepatitis C virus (anti-HCV). All HBsAg-positive sera were tested for the hepatitis B e antigen (HBeAg) and antibody (anti-HBe). The presence of antibodies to the hepatitis D virus was determined in most of the anti-Hbs-positive sera, and some of the HBsAg-positive sera. In addition to these, previous infection with syphilis, measles, HIV 1/2 and HTLV were determined by looking for the specific antibodies. We found HBsAg in 14.2% (20/141), anti-HBs in 93.6% (132/141), anti-HBs in 52.2% (73/140), anti-HCV in 7.9% (11/139, measles antibody in 99.3% (139/140), antibody to Treponema pallidum in 13.4% (18/134), antibody to HTLV-1 in 10.9% (15/138) and antibody to HIV-1 in 0.7% (1/140) of the sera tested.

Adolescent↗

An investigation of genital ulcers in Jackson, Mississippi, with use of a multiplex polymerase chain reaction assay: high prevalence of chancroid and human immunodeficiency virus infection.

In 1994, an apparent outbreak of atypical genital ulcers was noted by clinicians at the sexually transmitted disease clinic in Jackson, Mississippi. Of 143 patients with ulcers tested with a multiplex polymerase chain reaction (PCR) assay, 56 (39%) were positive for Haemophilus ducreyi, 44 (31%) for herpes simplex virus, and 27 (19%) for Treponema pallidum; 12 (8%) were positive for > 1 organism. Of 136 patients tested for human immunodeficiency virus (HIV) by serology, 14 (10%) were HIV-seropositive, compared with none of 200 patients without ulcers (P < .001). HIV-1 DNA was detected by PCR in ulcers of 6 (50%) of 12 HIV-positive patients. Multivariate analysis indicated that men with chancroid were significantly more likely than male patients without ulcers to report sex with a crack cocaine user, exchange of money or drugs for sex, and multiple sex partners. The strong association between genital ulcers and HIV infection in this population highlights the urgency of preventing genital ulcers in the southern United States.

Chancroid↗

Membrane topology and cellular location of the Treponema pallidum glycerophosphodiester phosphodiesterase (GlpQ) ortholog.

Recent reports that isolated Treponema pallidum outer membranes contain an ortholog for glycerophosphodiester phosphodiesterase (GlpQ) (D. V. Shevchenko, D. R. Akins, E. J. Robinson, M. Li, O. V. Shevchenko, and J. D. Radolf, Infect. Immun. 65:4179-4189, 1997) and that this protein is a potential opsonic target for T. pallidum (C. E. Stebeck, J. M. Shaffer, T. W. Arroll, S. A. Lukehart, and W. C. Van Voorhis, FEMS Microbiol. Lett. 154:303-310, 1997) prompted a more detailed investigation of its physicochemical properties and cellular location. [14C]palmitate radiolabeling studies of a GlpQ-alkaline phosphatase fusion expressed in Escherichia coli confirmed the prediction from DNA sequencing that the protein is lipid modified. Studies using Triton X-114 phase partitioning revealed that the protein's amphiphilicity is due to lipid modification and that a substantial portion of the polypeptide is associated with the T. pallidum peptidoglycan sacculus. Three different approaches, i.e., (i) proteinase K treatment of intact treponemes, (ii) indirect immunofluorescence analysis of treponemes encapsulated in agarose beads, and (iii) opsonophagocytosis of treponemes incubated with antiserum against recombinant GlpQ by rabbit peritoneal macrophages, confirmed that GlpQ is entirely subsurface in T. pallidum. Moreover, rabbits hyperimmunized with GlpQ were not protected against intradermal challenge with virulent treponemes. Circular dichroism spectroscopy confirmed that the recombinant form of the polypeptide lacked discernible evidence of denaturation. Finally, GlpQ was not radiolabeled when T. pallidum outer membranes were incubated with 3-(trifluoromethyl)-3-(m-[125I]iodophenyl)-diazarene, a photoactivatable, lipophilic probe which promiscuously labels both proteins and lipids within phospholipid bilayers. Taken as a whole, these studies indicate that the T. pallidum GlpQ ortholog is a periplasmic protein associated predominantly with the spirochete's peptidoglycan-cytoplasmic membrane complex.

Animals↗

Failure of penicillin treatment of yaws on Karkar Island, Papua New Guinea.

The endemic treponematosis yaws remains a significant cause of morbidity in many tropical countries, despite mass treatment campaigns to eradicate it. An outbreak of yaws in Marup village on Karkar Island, Papua New Guinea in 1988 provided an opportunity to monitor the outcome of treatment with penicillin over an extended period. Thirty-nine children with clinical yaws (6% of 632 examined) were monitored clinically and serologically, for nearly two years after mass treatment of all villagers with the World Health Organization recommended dosages of benzathine penicillin. Lesions resolved within one month of treatment in all but four (10%) children, three of whom were initially successfully retreated. Before treatment, the Venereal Disease Research Laboratory (VDRL) test result was reactive in 67% of the children and treponema-specific IgM antibody test results were reactive in 41%. Within six months of treatment, of those reactive, the VDRL titer decreased significantly in 25 (96%) of 26 and IgM antibody test results became negative in 13 (81%) of 16 children. However, by the end of follow-up, 11 (28%) of the 39 children had developed clinical and/or serologic evidence of relapse. In these children, response to further treatment was slow and, in three, evidence of active infection persisted or recurred, despite repeated courses. Exogenous reinfection was unlikely in this isolated community, in which the occurrence of yaws was closely monitored after universal treatment. Treatment failure was most likely to have been due to reduced susceptibility to penicillin of Treponema pallidum subsp. pertenue.

Adolescent↗

Epidemiology of adult sexually transmitted disease agents in children being evaluated for sexual abuse.

This prospective study describes the epidemiology of adult sexually transmitted disease agents in 1538 children ages 1 to 12 years being evaluated for possible sexual abuse. Infections with these agents were related to the presence or absence of a history of sexual contact. Neisseria gonorrhoeae (GC) was found in 2.8% (41 of 1469); human papillomavirus presenting as condyloma acuminata, 1.8%; Chlamydia trachomatis, 1.2% (17 of 1473); Treponema pallidum (syphilis), 0.1% (1 of 1263); and herpes simplex virus, 0.1%. Overall a history of sexual contact was present in 83% of children with N. gonorrhoeae; condyloma acuminata, 43%; Chlamydia trachomatis, 94%; syphilis, 0%; and herpes simplex virus, 50%. Selected vaginal discharges were examined for Trichomonas vaginalis and bacterial vaginosis. In children comprehending questions regarding sexual contact (i.e. were "verbal"), 89% with N. gonorrhoeae, 100% with Chlamydia trachomatis and 63% with condyloma acuminata had a history of sexual contact, indicating that in "verbal" children any infection with N. gonorrhoeae or C. trachomatis was highly associated with sexual contact.

Child↗

Enhanced retention of motility and virulence of Treponema pallidum (Nichols strain) in vitro by the addition of gelatin to anaerobic medium.

The effect of gelatin on the survival of Treponema pallidum (Nichols strain) in anaerobic maintenance medium was investigated by monitoring of the retention of treponemal motility and virulence. Motility of T. pallidum was determined by dark-field microscopy, and virulence of T. pallidum was measured by observation of the latent periods of infection after inoculation of rabbits with virulent organisms. Gelatin concentrations of zero, 3%, 5%, and 15% were tested, and optimal retention of treponemal motility and virulence was observed at 5% and 10% gelatin.

Animals↗

Multiple alleles of Treponema pallidum repeat gene D in Treponema pallidum isolates.

Two new tprD alleles have been identified in Treponema pallidum: tprD2 is found in 7 of 12 T. pallidum subsp. pallidum isolates and 7 of 8 non-pallidum isolates, and tprD3 is found in one T. pallidum subsp. pertenue isolate. Antibodies against TprD2 are found in persons with syphilis, demonstrating that tprD2 is expressed during infection.

Alleles↗

Production of murine monoclonal antibodies to the major axial filament polypeptide of Treponema pallidum.

A suitable immunization protocol for the stimulation of a murine antibody response to the axial filament polypeptides of Treponema pallidum was established. A range of monoclonal antibodies (Mabs) specific for different epitopes of the major axial filament polypeptide (37 kDa) was generated which demonstrated diversity in their ability to react with other treponemal species. Immunogold electron microscopy located the 37 kDa antigen on the surface of the axial filament structure. The early appearance of specific antibody to this polypeptide in infected man and rabbit indicates that such Mabs are potentially useful for the diagnosis of early syphilis.

Animals↗